WO2003050274A2 - Production de cristaux a partir de bouillon de fermentation - Google Patents

Production de cristaux a partir de bouillon de fermentation Download PDF

Info

Publication number
WO2003050274A2
WO2003050274A2 PCT/DK2002/000821 DK0200821W WO03050274A2 WO 2003050274 A2 WO2003050274 A2 WO 2003050274A2 DK 0200821 W DK0200821 W DK 0200821W WO 03050274 A2 WO03050274 A2 WO 03050274A2
Authority
WO
WIPO (PCT)
Prior art keywords
fermentation broth
polymer
metabolite
fusarium
crystalline
Prior art date
Application number
PCT/DK2002/000821
Other languages
English (en)
Other versions
WO2003050274A3 (fr
Inventor
Benny Nielsen
Anders Rancke-Madsen
Martin Troen JØRGENSEN
Original Assignee
Novozymes A/S
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Novozymes A/S filed Critical Novozymes A/S
Priority to AT02781173T priority Critical patent/ATE547518T1/de
Priority to EP02781173A priority patent/EP1456361B1/fr
Priority to DK02781173.6T priority patent/DK1456361T3/da
Priority to AU2002349300A priority patent/AU2002349300A1/en
Publication of WO2003050274A2 publication Critical patent/WO2003050274A2/fr
Publication of WO2003050274A3 publication Critical patent/WO2003050274A3/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2405—Glucanases
    • C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
    • C12N9/2411—Amylases
    • C12N9/2414—Alpha-amylase (3.2.1.1.)
    • C12N9/2417—Alpha-amylase (3.2.1.1.) from microbiological source
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14—Extraction; Separation; Purification
    • C07K1/30—Extraction; Separation; Purification by precipitation
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14—Extraction; Separation; Purification
    • C07K1/30—Extraction; Separation; Purification by precipitation
    • C07K1/306—Extraction; Separation; Purification by precipitation by crystallization
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
    • C12N9/54—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea bacteria being Bacillus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione

Definitions

  • the present invention relates to a simple and effective method for obtaining a crystalline and/or amorphous metabolite suspension from a fermentation broth.
  • the present invention provides a simple and effective method for producing a crystalline and/or amorphous metabolite suspension from a fermentation broth.
  • the method of the invention may be applied to an untreated fermentation broth or to a fermentation broth that has first been subjected to, e.g., a pH adjustment, a temperature adjustment, and/or a water dilution.
  • Metabolites of interest may be an antibiotic such as penicillin or cephalosporin, or a commodity chemical such as citric acid.
  • the metabolite may also be a protein, e.g. a therapeutic protein such as insulin or an enzyme.
  • the enzyme may be a hydrolase, a transferase, a lyase, an isomerase, or a ligase.
  • the method is applied to proteases, lipases, amylases, cellulases, and oxidoreductases.
  • proteases include those of animal, vegetable or microbial origin. Microbial origin is preferred. Chemically modified or protein engineered mutants are included.
  • the protease may be a serine protease or a metallo protease, preferably an alkaline microbial protease or a trypsin-like protease.
  • alkaline proteases are subtilisins, especially those derived from Bacillus, e.g., subtilisin Novo, subtilisin Carlsberg, subtilisin 309, subtilisin 147 and subtilisin 168 (described in WO 89/06279).
  • trypsin-like proteases are trypsin (e.g. of porcine or bovine origin) and the Fusarium protease described in 5 WO 89/06270 and WO 94/25583.
  • Suitable lipases include those of bacterial or fungal origin. Chemically modified or protein engineered mutants are included. Examples of useful lipases include lipases from Humicola (synonym Thermomyces), e.g. from H.
  • lanuginosa T. lanuginosus
  • a Pseudomonas lipase e.g. from P. alcaligenes or P. pseudoalcaligenes (EP is 218 272), P. cepacia (EP 331 376), P. stutzeri (GB 1 ,372,034), P. fluorescens, Pseudomonas sp. strain SD 705 (WO 95/06720 and WO 96/27002), P. wisconsinensis (WO 96/12012), a Bacillus lipase, e.g.
  • B. subtilis (Dartois et al. (1993), Biochemica et Biophysica Acta, 1131 , 253-360), B. stearothermophilus (JP 64/744992) or B. pumilus (WO 91/16422).
  • lipase variants such as those described in WO 92/05249, WO
  • Amylases include those of bacterial or fungal origin. Chemically modified or protein engineered mutants are included. Amylases
  • alpha-amylases obtained from Bacillus, e.g. a special strain of B. licheniformis, described in more detail in GB 1 ,296,839.
  • amylases examples include the variants described in WO 94/02597, WO 94/18314, WO 96/23873, and WO 97/43424, especially the variants with substitutions in one or more of the following positions: 15, 23, 105, 106, 124, 128, 133, 154, 156, 181 , 188,
  • Suitable cellulases include those of bacterial or fungal origin. Chemically modified or protein engineered mutants are included. Suitable cellulases include cellulases from the genera Bacillus, Pseudomonas, Humicola, Fusarium, Thielavia, Acremonium, e.g. the fungal cellulases produced from Humicola insolens, Myceliophthora thermophila and Fusarium oxysporum disclosed in US 4,435,307, US 5,648,263, US 5,691 ,178, US 5,776,757 and WO 89/09259.
  • cellulases are the alkaline or neutral cellulases having colour care benefits.
  • Examples of such cellulases are cellulases described in EP 0 495 257, EP 0
  • Oxidoreductases that may be treated according to the invention include peroxidases (EC 1.11.1.7), and oxidases such as laccases, and catalases (EC 1.11.1.6).
  • Peroxidases Preferably, the peroxidase employed in the method of the invention is producible by microorganisms such as fungi or bacteria.
  • Coprinus peroxidase is preferred, in particular a C. macrorhizus or C. cinereus peroxidase, or a variant thereof.
  • laccases and laccase related enzymes contemplate any laccase enzyme comprised by the enzyme classification (EC 1.10.3.2), any chatechol oxidase enzyme comprised by the enzyme classification (EC 1.10.3.1), any bilirubin oxidase enzyme comprised by the enzyme classification (EC 1.3.3.5) or any monophenol monooxygenase enzyme comprised by the enzyme classification (EC 1.14.18.1).
  • the microbial laccase enzyme may be derived from bacteria or fungi (including filamentous fungi and yeasts) and suitable examples include a laccase derivable from a strain of Aspergill us, Neurospora, e.g., N. crassa, Podospora, Botrytis, Collybia, Fomes, Lentinus, Pleurotus, Trametes, e.g., T. villosa and T. versicolor, Rhizoctonia, e.g., R. solani, Coprinus, e.g. C. plicatilis and C. cinereus, Psatyrella, Myceliophthora, e.g. M.
  • Neurospora e.g., N. crassa, Podospora, Botrytis, Collybia, Fomes, Lentinus, Pleurotus, Trametes, e.g., T. villosa and T. versicolor
  • thermophila Schytalidi- um
  • Polyporus e.g., P. pinsitus
  • Phlebia e.g., P. radita
  • Co olus e.g., C. hirsutus (JP 2-238885).
  • the fermentation broth according to the invention may be obtained from any microorganism of any genus known in the art.
  • the metabolite of interest may be obtained from a bacterial or a fungal source.
  • the metabolite of interest may be obtained from a gram positive bacterium such as a Bacillus strain, e.g., Bacillus alkalophilus, Bacillus amyloliquefaciens, Bacillus brevis, Bacillus circulans, Bacillus coagulans, Bacillus lautus, Bacillus lentus, Bacillus clausii, Bacillus licheniformis, Bacillus megaterium, Bacillus stearothermophilus, Bacillus subtilis, or Bacillus thuringiensis; or a Streptomyces strain, e.g., Streptomyces lividans or Streptomyces murinus; or from a gram negative bacterium, e.g., E. coli or Pseudomonas sp.
  • a Bacillus strain e.g., Bacillus alkalophil
  • the metabolite of interest may be obtained from a fungal source, e.g. from a yeast strain such as a Candida, Kluyveromyces, Pichia, Saccharomyces, Schizosaccharomyces, or Yarrowia strain, e.g., Saccharomyces carlsbergensis, Saccharomyces cerevisiae, Saccharomyces diastaticus, Saccharomyces douglasii, Saccharomyces kluyveri, Saccharomyces norbensis or Saccharomyces oviformis strain.
  • yeast strain such as a Candida, Kluyveromyces, Pichia, Saccharomyces, Schizosaccharomyces, or Yarrowia strain, e.g., Saccharomyces carlsbergensis, Saccharomyces cerevisiae, Saccharomyces diastaticus, Saccharomyces douglasii, Saccharomyces kluyver
  • the metabolite of interest may be obtained from a filamentous fungal strain such as an Acremonium, Aspergillus, Aureobasidium, Cryptococcus, Filibasidium, Fusarium, Humicola, Magnaporthe, Mucor, Myceliophthora, Neocallimastix, Neurospora, Paecilomyces, Penicillium, Piromyces, Schizophyllum, Talaromyces, Thermoascus, Thielavia, Tolypocladium, or Trichoderma strain, in particular the metabolite of interest may be obtained from an Aspergillus aculeatus, Aspergillus awamori, Aspergillus foetidus, Aspergillus japonicus, Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae, Fusarium bactridioides, Fusarium cerealis, Fusarium bactridio
  • the term "obtained from” as used herein in connection with a given source shall mean that the metabolite of interest is produced by the source or by a cell in which a gene from the source has been inserted.
  • the microbial strain may be fermented by any method known in the art.
  • the fermentation medium may be a complex medium comprising complex nitrogen and/or carbon sources, such as soybean meal, cotton seed meal, corn steep liquor, yeast extract, casein hydrolysate, molasses, and the like.
  • the fermentation medium may be a chemically defined media, e.g. as defined in WO 98/37179.
  • the fermentation may be performed as a fed-batch, a repeated fed-batch or a continuous fermentation process.
  • a useful coagulant may be a salt such as one selected from the group consisting of the group I metal salts, the group II metal salts, and the group III metal salts, or mixtures thereof; in particular Ca-, Mg- and Al-salts.
  • Preferred salts are ammonium, phosphate, sulfate, carbonate, and citrate salts.
  • Halogenide salts, formiates and acetates may also be applicable, especially chloride salts such as calcium chloride.
  • Useful salt concentrations will be in the range of 0.1-40% (w/w); preferably in the range of 0.2-20% (w/w); more preferably in the range of 0.3-6% (w/w).
  • another useful coagulant is a short chained polymer, in particular a cationic polymer with a molecular weight in the range of from 20 Daltons to 500000 Daltons, such as a tertiary or a quaternary polyamine, e.g. C521 obtainable from Cytec Industries, PoIy-DADMAC's (Di-allyl Dimethyl Ammonium Chlorid), e.g. C591 , or Aluminimu sources such as polyaluminumchlorohydrate: AI2(OH)5CI, e.g. GC850 obtainable from Gulbrandsen.
  • AI2(OH)5CI e.g. GC850 obtainable from Gulbrandsen.
  • Useful short chained polymer concentrations will typically be in the range of 0.1- 25% (w/w); preferably in the range of 0.2-20% (w/w); more preferably in the range of 0.3-15% (w/w).
  • coagulant e.g. a salt and one or more short chained polymers.
  • a useful flocculant may be an inorganic and/or organic polymer which may be cationic, anionic or non-ionic.
  • a useful cationic polymer is a polyamine, and a useful anionic polymer is a polyacrylamid.
  • Useful polymer concentrations will be in the range of 0.01-1.0% (w/w); preferably in the range of 0.05-0.5% (w/w).
  • a useful separation equipment is any design of a two- phase centrifuge, especially a continuous sludge decharging centrifuge, a decanter or a cyclone.
  • a flocculatant in particular an anionic polymer
  • coagulate and/or flocculate the metabolite fermentation broth so that the crystalline and/or amorphous metabolites are in the right separation zone and thus can be separated in e.g. a centrifuge process into a biomass fraction (with a very low metabolite concentration), the crystalline and/or amorphous metabolite fraction (with a high metabolite concentration) and the supernatant fraction (with a very low metabolite concentration).
  • the coagulation and/or the flocculation has the effect that the crystalline and/or amorphous metabolites are not incorporated in the flocks, so after the biomass has been separated the crystalline and/or amorphous metabolites may be further concentrated in order to achieve the wanted yield.
  • the suspension achieved according to the invention may be further purified in a variety of ways such as by using grinding, sieving, drying, filtration, centrifugation, re- crystallisation, chromatographic methods, adsorption processes and/or two-phase extraction. •
  • the invention is further illustrated in the following examples which are not intended to be in any way limiting to the scope of the invention as claimed.
  • a fermentation broth containing a mutated subtilisin protease obtained from a Bacillus sp. disclosed in WO 91/00345 was subjected to the method of the invention.
  • the protease fermentation broth was run in production scale based on the following coagulation/flocculation recipe:
  • the trial was made on an entire protease batch using one Westfalia SB60 centrifuge.
  • the feed contained 17-20% wet sludge volume.
  • the centrifuge used was equipped with a nozzle bowl and a standardized sludge flow of 4 m3/hr.
  • the trial ran 21 hours where the feed flow was step-wise increased from 7.5 m3/h to 9.5 m3/hr.
  • an anionic polymer was added at feed flows above 7.5 m3/hr.
  • the anionic polymer solution used was 0.15% (w/w) (polyacrylamid).
  • the percent wet crystal volume was measured in the centrate during the trial and used as yield indicator.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Molecular Biology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Microbiology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Analytical Chemistry (AREA)
  • Biophysics (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Mushroom Cultivation (AREA)

Abstract

Procédé de production de suspension de métabolites cristalline et/ou amorphe à partir d'un bouillon de fermentation, qui comporte les étapes consistant à traiter le bouillon de fermentation à l'aide d'un ou de plusieurs coagulants et/ou d'un ou de plusieurs floculants ; et à séparer la biomasse du bouillon de fermentation coagulé et/ou floculé à l'aide d'un équipement de séparation, ce qui permet d'obtenir une suspension de métabolites cristalline et/ou amorphe.
PCT/DK2002/000821 2001-12-11 2002-12-04 Production de cristaux a partir de bouillon de fermentation WO2003050274A2 (fr)

Priority Applications (4)

Application Number Priority Date Filing Date Title
AT02781173T ATE547518T1 (de) 2001-12-11 2002-12-04 Verfahren zur sammlung von kristallinen partikeln aus fermentationsbrühe
EP02781173A EP1456361B1 (fr) 2001-12-11 2002-12-04 Production de cristaux a partir de bouillon de fermentation
DK02781173.6T DK1456361T3 (da) 2001-12-11 2002-12-04 Fremgangsmåde til opsamling af krystallinske partikler fra fermenteringsmedium
AU2002349300A AU2002349300A1 (en) 2001-12-11 2002-12-04 Process for harvesting crystalline particles from fermentation broth

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DKPA200101847 2001-12-11
DKPA200101847 2001-12-11

Publications (2)

Publication Number Publication Date
WO2003050274A2 true WO2003050274A2 (fr) 2003-06-19
WO2003050274A3 WO2003050274A3 (fr) 2003-12-11

Family

ID=8160891

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/DK2002/000821 WO2003050274A2 (fr) 2001-12-11 2002-12-04 Production de cristaux a partir de bouillon de fermentation

Country Status (6)

Country Link
EP (1) EP1456361B1 (fr)
CN (1) CN100487114C (fr)
AT (1) ATE547518T1 (fr)
AU (1) AU2002349300A1 (fr)
DK (1) DK1456361T3 (fr)
WO (1) WO2003050274A2 (fr)

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005042758A1 (fr) * 2003-10-31 2005-05-12 Novozymes A/S Procede de floculation d'un bouillon de fermentation comprenant un champignon
WO2005063794A1 (fr) * 2003-12-30 2005-07-14 Bharat Biotech International Limited Procede de preparation et de purification de proteines de recombinaison
EP2125867A1 (fr) * 2006-12-22 2009-12-02 Bayer Technology Services GmbH Dispositif et procédé de précipitation de peptides
WO2009156073A1 (fr) * 2008-06-23 2009-12-30 Bayer Technology Services Gmbh Procédé de séparation sélective de peptides et de protéines par cristallisation
EP2196539A1 (fr) * 2008-12-15 2010-06-16 Technische Universiteit Delft Procédé pour production continue biologique des lipides, glucides ou leurs mélanges
CN102838552A (zh) * 2012-09-21 2012-12-26 江南大学 采用复合絮凝法生产高纯度吩嗪-1-羧酸的方法
US8889395B2 (en) 2010-03-30 2014-11-18 Novozymes A/S Crystal metabolite recovery
EP2914611B1 (fr) 2012-11-01 2018-08-29 Novozymes A/S Procédé d'élimination d'adn
WO2018185048A1 (fr) 2017-04-03 2018-10-11 Novozymes A/S Procédé de récupération
WO2019160862A1 (fr) * 2018-02-13 2019-08-22 Lygos, Inc. Procédé de préparation de dérivés diesters de l'acide malonique
WO2020074517A1 (fr) 2018-10-10 2020-04-16 Novozymes A/S Variants d'inhibiteur de chymotrypsine et utilisation associée
WO2024032881A1 (fr) * 2022-08-10 2024-02-15 Arla Foods Amba Cristallisation d'une protéine pouvant cristalliser dans des conditions de salage à l'aide de multiples charges protéiques

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106146610B (zh) * 2016-09-22 2018-01-23 济南大学 一种采用絮凝法制备谷胱甘肽亚铜盐的方法

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0574050A1 (fr) 1992-05-19 1993-12-15 Gist-Brocades N.V. Séparation et purification du produit de fermentation à grande échelle

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4659667A (en) * 1985-02-26 1987-04-21 Miles Laboratories, Inc. Process to recover crystalline enzymes and crystalline enzymes produced thereby
JP2958789B2 (ja) * 1990-01-22 1999-10-06 味の素株式会社 アミノ酸・核酸およびその誘導体の精製方法
DK83093D0 (da) * 1993-07-09 1993-07-09 Novo Nordisk As Fremgangsmaade
CN1114613C (zh) * 1995-06-02 2003-07-16 诺沃奇梅兹有限公司 蛋白质溶液的铝/铁处理和膜浓缩方法

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0574050A1 (fr) 1992-05-19 1993-12-15 Gist-Brocades N.V. Séparation et purification du produit de fermentation à grande échelle

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005042758A1 (fr) * 2003-10-31 2005-05-12 Novozymes A/S Procede de floculation d'un bouillon de fermentation comprenant un champignon
WO2005063794A1 (fr) * 2003-12-30 2005-07-14 Bharat Biotech International Limited Procede de preparation et de purification de proteines de recombinaison
US20070154880A1 (en) * 2003-12-30 2007-07-05 Ella Krishna M Process for the preparation and purification of recombinant proteins
US8956812B2 (en) * 2003-12-30 2015-02-17 Bharat Biotech International Limited Process for the preparation and purification of recombinant proteins
EP2125867A1 (fr) * 2006-12-22 2009-12-02 Bayer Technology Services GmbH Dispositif et procédé de précipitation de peptides
EP2125867A4 (fr) * 2006-12-22 2011-05-11 Bayer Technology Services Gmbh Dispositif et procédé de précipitation de peptides
WO2009156073A1 (fr) * 2008-06-23 2009-12-30 Bayer Technology Services Gmbh Procédé de séparation sélective de peptides et de protéines par cristallisation
EP2196539A1 (fr) * 2008-12-15 2010-06-16 Technische Universiteit Delft Procédé pour production continue biologique des lipides, glucides ou leurs mélanges
US8889395B2 (en) 2010-03-30 2014-11-18 Novozymes A/S Crystal metabolite recovery
CN102838552A (zh) * 2012-09-21 2012-12-26 江南大学 采用复合絮凝法生产高纯度吩嗪-1-羧酸的方法
EP2914611B1 (fr) 2012-11-01 2018-08-29 Novozymes A/S Procédé d'élimination d'adn
WO2018185048A1 (fr) 2017-04-03 2018-10-11 Novozymes A/S Procédé de récupération
WO2019160862A1 (fr) * 2018-02-13 2019-08-22 Lygos, Inc. Procédé de préparation de dérivés diesters de l'acide malonique
US11597951B2 (en) 2018-02-13 2023-03-07 Lygos, Inc. Method for preparation of diester derivatives of malonic acid
WO2020074517A1 (fr) 2018-10-10 2020-04-16 Novozymes A/S Variants d'inhibiteur de chymotrypsine et utilisation associée
WO2024032881A1 (fr) * 2022-08-10 2024-02-15 Arla Foods Amba Cristallisation d'une protéine pouvant cristalliser dans des conditions de salage à l'aide de multiples charges protéiques

Also Published As

Publication number Publication date
CN100487114C (zh) 2009-05-13
EP1456361A2 (fr) 2004-09-15
WO2003050274A3 (fr) 2003-12-11
CN1599795A (zh) 2005-03-23
AU2002349300A1 (en) 2003-06-23
EP1456361B1 (fr) 2012-02-29
AU2002349300A8 (en) 2003-06-23
ATE547518T1 (de) 2012-03-15
DK1456361T3 (da) 2012-06-11

Similar Documents

Publication Publication Date Title
Aunstrup Production, isolation and economics of extracellular enzymes
WO2003050274A2 (fr) Production de cristaux a partir de bouillon de fermentation
US20240191175A1 (en) Method of purifying a protein from fermentation solids under desorbing conditions
EP0828753B1 (fr) Traitement d'une solution proteique avec un compose d'al/fe, suivi d'une concentration membranaire
US20070015254A1 (en) Crystal harvest from fermentation broth
EP1520011A2 (fr) Sterilisation d'une culture de fermentation contenant source n hydrolysee
WO2015144936A1 (fr) Resolubilisation de cristaux de protéines à faible ph
WO2018185048A1 (fr) Procédé de récupération
EP0894091A1 (fr) Procede de purification d'une proteine presente dans une solution contenant des proteines
US20120122183A1 (en) Flocculation with divalent salt and phosphate
US6031082A (en) Increased yields of a crystallized protein by using a solid adsorption material
US9347080B2 (en) Use of browned glucose as a feed substrate
DK2552934T3 (en) Krystalmetabolit-extraction
WO2004003187A2 (fr) Adjonction de monopropylene glycol a une fermentation
US20100173286A1 (en) Mutant Cells Suitable for Recombinant Polypeptide Production
US7019120B2 (en) Cloud-point extraction of enzymes and polypeptides from a fermentation broth using a non-ionic surfactant
US20040033567A1 (en) Hydrolysed N-source
EP2213723A1 (fr) Isomaltose pour la fermentation fongique
WO2020173473A1 (fr) Polypeptides à domaine chap et leur utilisation pour traiter des boues
CN113993878A (zh) 用二价阳离子从发酵液中回收蛋白的方法
CN111032856A (zh) 基于pH的FCA控制的用途

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ OM PH PL PT RO RU SC SD SE SG SK SL TJ TM TN TR TT TZ UA UG UZ VC VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR IE IT LU MC NL PT SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
WWE Wipo information: entry into national phase

Ref document number: 2002781173

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 20028243110

Country of ref document: CN

WWP Wipo information: published in national office

Ref document number: 2002781173

Country of ref document: EP

NENP Non-entry into the national phase

Ref country code: JP

WWW Wipo information: withdrawn in national office

Country of ref document: JP